Stable Isotope Ratio Analysis Research Articles

A biogeochemical approach to examining sub-adult diet and the weaning process at Neolithic Tepecik-Çiftlik, Türkiye

Tepecik-Çiftlik, a Neolithic settlement located in the Cappadocia region of Central Anatolia, within the boundaries of Niğde province in modern Türkiye is crucial for helping us to better understand social behaviour and human mobility in the Neolithic, as well as relations between different groups. To investigate dietary habits, subsistence practices, and intra-societal variations stable carbon and nitrogen isotope ratio analysis has been conducted on the Neolithic human and faunal population. Further to this, 21 sub-adults were also sampled for stable carbon and nitrogen isotope ratio analysis to examine sub-adult diet, complementary feeding, and the cessation of breastfeeding as part of the weaning process. This isotopic investigation into the weaning process is complemented by a previously conducted study using Sr/Ca ratios from bone apatite of the same population. The δ13C values of the sampled sub-adults range from −19.8 ‰ to −19.0 ‰ and the δ15N values range from 8.0 ‰ to 15.2 ‰. The isotopic data suggest a very early onset of complementary feeding (ca. 0.2 years of age) and a brief breastfeeding period, relative to other prehistoric populations. The cessation of breastfeeding and this very brief weaning process was likely completed by ca. 1–1.5 years old. This early commencement of complementary feeding was most probably a deliberate social and cultural choice, supported by the presence of bowls and feeding spoons in the graves of very young babies and infants. However, this cultural choice may also have affected the mortality profile of the Neolithic population.

Egg authentication under seasonal variation using stable isotope analysis combined with machine learning classification

In the European Union (EU) eggs from laying hens are sold in different husbandry systems: enriched cage, barn, free-range and organic. An analytical distinction between the husbandry systems is of interest for the food control authorities, as fraudsters can make high profits by mislabelling the husbandry system of the eggs. In this study, a total of 180 hen's egg samples labelled to originate from barn, free-range and organic hens were taken from food retailers in Germany at monthly intervals over a period of 15 months. The eggs were analysed using stable isotope ratio analysis (IRMS) in order to identify potential influences of different husbandry methods. The analysis on a monthly basis showed that the feeding of the laying hens varied over the year, which makes it difficult to make a clear classification according to the type of husbandry. Temporary events such as the officially ordered poultry quarantine due to the avian influenza, had minor influence on the differentiation of the husbandry types. Various statistical models including linear discriminant analysis (LDA), random forest (RF) and multinomial logistic regression (Multinomial) based on a multi-element approach (δ13C, δ15N, δ34S) were used to classify the eggs according to the three types of husbandry. On average, the models achieved a correct classification of approximately 70%. The reduction of the prediction model to two classes (organic vs. conventional eggs) led to a correct classification of almost 80% across all samples. Additionally, significant differences in the mean values of the sulphur isotopes of the organic eggs showed a regional differentiation between Bavaria (southern Germany) and Schleswig-Holstein (northern Germany).

Tracking mercury sources in the Wabigoon River: Use of stable mercury isotopes in bioindicator organisms

Mercury concentrations remain elevated in sediments and biota of the Wabigoon River downstream from Dryden, Ontario, the home of a former chlor-alkali plant. Understanding the current extent and severity of mercury contamination downstream of this industrial legacy site is of great importance in managing the mercury contamination within the traditional territory of Asubpeeschoseewagong Anishinabek (Grassy Narrows First Nation), located downstream of Dryden. The objective of this study was to use mercury stable isotope ratio analysis to distinguish between legacy mercury from the former chlor-alkali plant and mercury from geogenic sources. Mercury concentrations in surface sediments and biota downstream of the historical source of mercury discharge are elevated relative to the chosen reference location, Wabigoon Lake (WL). Mean sediment mercury levels were as high as 3.27 μg/g at the hydroelectric dam location compared to 0.05–0.10 μg/g at Wabigoon Lake sediments. Isotope ratios in aquatic biota and sediments collected from within the system were distinct from Wabigoon Lake, indicating that anthropogenic mercury contamination is distinguishable from geogenic mercury. Average δ202Hg values of −2.46 ± 0.41 observed in sediments of WL were consistently more negative compared to downstream values, which varied from −1.34 to 0.30 ‰. Young-of-the-year Yellow Perch and Hexagenia were found to have significantly more positive δ202Hg values downstream from Wabigoon Lake.

Pretreatment method for oxygen stable isotope ratio analysis of the sugar-rich fraction in fruit juice via isotope ratio mass spectrometry.

The oxygen stable isotope ratio (δ18O) of the sugar-rich fraction of fruit juice is important as a tracer of the geographical origin of raw material. This study sought to minimize the inter-day variation of δ18O attributable to the influence of water to accurately monitor geographical origin labeling. Two drying devices (freeze dryer and vacuum oven) were compared. Then, two humidity levels (normal and low humidity) at which the samples were placed after drying were compared. The low-humidity environment was constructed using a glove bag and pure argon gas. δ18O was measured using thermal conversion elemental analyzer/isotope ratio mass spectrometry. Improvements were made to the measurement method based on aforementioned analyses results, and the performance of the initial and improved methods was compared. δ18O of juice dried in a vacuum oven was 3.30‰ lower than that of juice dried in a freeze dryer. Moreover, δ18O of juice samples exposed to normal humidity was 3.74‰ lower than that of samples exposed to low humidity. The combined inter-day and intra-day standard deviation was reduced from 1.20‰ in the initial method to 0.42‰ in the improved method. This study describes a pretreatment method for δ18O measurement in the sugar-rich fraction of fruit juice with less inter-day variation, and it will be useful for monitoring geographical origin labeling.

Bioarchaeological Perspectives on Late Antiquity in Dalmatia: Paleogenetic, Dietary, and Population Studies of the Hvar—Radošević burial site

Late Antiquity Dalmatia was a time and place of political unrest in the Roman Empire that influenced the lives of those in that region. The Late Antique burial site of Hvar – Radošević, spanning the 3rd to 5th centuries CE, is located on the Croatian Dalmatian island of Hvar. Given the time frame and location on a busy marine trade route, the study of this burial site offers us a glimpse into the lives of the Late Antique population living on this island. It comprises 33 individuals, with 17 buried within a confined grave tomb and the remaining individuals buried in separate locations in the tomb's proximity. The study aims to provide a new perspective on the lives of people on the island during those times by studying ancestry, population structure, possible differences within the buried population, dietary habits, and general health. The genetic analysis of the ancestral origins of the individuals buried at Hvar – Radošević revealed a diverse population reflective of the era's genetic variability. The identification of genetic outliers suggests a range of ancestries from distinct regions of the Roman Empire, possibly linked to trade routes associated with the Late Antique port in ancient Hvar. Stable isotope ratio analysis (δ13C and δ15N) indicated a diet mainly consisting of C3 plants, with minimal consumption of marine foods. High childhood mortality rates, physiological stress markers, and dental diseases suggest a low quality of life in the population. Assessment of kinship and dietary patterns revealed no discernible distinctions between individuals buried within the tomb and those buried outside, indicative of an absence of differential burial practices based on social status and familial ties among this specific buried population.

Use of stable isotope ratio analysis to investigate the biology and clinical significance of seal parasites.

Stranded seals are often infected with a broad range of parasites, although whether they are the cause of significant morbidity or an incidental finding is usually unclear. In this study we used stable isotope ratio analysis, a method frequently used to investigate food webs, to explore the extent to which common seal parasites feed directly on host tissue and fluids or absorb host-derived metabolites, which in turn may give an indication of their potential impact on the host's health. The trophic discrimination factor Δ15N for the nasal mite, Halarachne halichoeri, was mostly positive, ranging between −0.015 and 3.2‰ (n = 6), while for the Acanthocephalan worm, Corynosoma strumosum and the anisakid nematode, Pseudoterranova decipiens, Δ15N ranged between −4.2 and −2.0‰ (n = 7), and between −6.7 and −0.8‰ (n = 5) respectively. In the case of the lungworm, Otostrongylus circumlitus, Δ15N measured between −5.6 and 0‰ for worms collected in the stomach (n = 5), between −1.1 and 0.2‰ for worms collected from the heart (n = 3), between −0.7 and 1.9‰ for worms situated in the lungs (n = 4). Based on Δ15N, parasites could be clearly divided into those that were on a higher trophic level than their host suggesting a predator–prey-like relationship, and those that were not. It is hypothesized that Δ15N may be indicative of the clinical significance of parasite–host associations.

CO2 conversion to CO by fluidized bed biomass gasification: Measuring CO2 utilization via stable carbon isotope ratios

Thermochemical conversion of CO2 in biomass gasification is a promising technology for utilizing CO2 as a feedstock to produce a CO-rich gas. Simultaneous decomposition reactions of biomass and various gas-solid and gas-gas reactions form the product gas in this process. The overlap in sub-processes makes it challenging to assess the conversion of feedstock CO2 with common methods like mass balancing. This work introduces stable carbon isotope ratio analysis (δ13C) to identify the sourcing of carbonaceous product gas components and determine the conversion of CO2. This methodology is applied to evaluate experiments conducted for one hour of continuous operation in a lab-scale fluidized bed gasifier. Softwood pellets and wood char are used as fuel, with Olivine as a bed material, a target heating temperature of 1000 °C and atmospheric pressure. Product gas with more than 80 vol% CO was generated when wood char was used as fuel. Stable carbon isotope measurements show that CO2 is converted at 48–93% in this process, underpinning the position of biomass CO2 gasification as carbon capture and utilization technology. These results were up to 25% higher than suggested by mass balancing, with higher discrepancies at lower CO2 conversions when using softwood as fuel. Therefore, stable carbon isotope ratio measurement can be a valuable tool for improving the process understanding of biomass CO2 gasification. The results can be used for carbon accounting and the technical development of gasifiers with high CO2 utilization efficiency.

Stable isotope insights into arthropod food chains and nitrogen cycling in a rehabilitated tailings chronosequence

Field studies in post-mining sites on epigeic invertebrate groups are scarce despite their importance in pedogenesis and ecosystem multifunctionality. This research investigated the diversity, abundance and succession of aboveground invertebrates in a rehabilitated Pb/Zn tailings chronosequence of 5, 20 and 35 years. The study also explored the trophic relationship of selected arthropods and characterized the nitrogen (N) cycle using stable isotope measurements. The abundance and species richness in most investigated groups and the dominance index Berger–Parker (BP) of aboveground invertebrates increased with rehabilitation age from 0.17 BP in early to 0.31 BP in late stage. Elemental and stable isotope ratio analysis showed that N and C soil content increased and the C/N ratio decreased with age, yet despite this increased N availability in the system, the maturing N-cycle used N efficiently. The lack of large N losses from the system despite substantial atmospheric deposition inputs was indicated by the fact that N isotope ratios (δ15N) in plants and animals became significantly more negative with rehabilitation age, −6.0 δ15N for plants, −5.0 δ15N for herbivores and 3.0 δ15N for carnivores. The length of the invertebrate food chain expanded by more than half a trophic level (2.7‰ δ15N) for top predator Coleoptera from early to late stage, probably reflecting more complex food webs including intra-guild predation in older communities. In conclusion, δ15N measurements in plants and animals provided novel insights into the N-cycle, accumulative N flows and the trophic position in post-mining sites. It is proposed that isotope ratio measurements could be used as easy-to-measure, integrating indicators of nutrient cycling and the soil food web complexity of rehabilitated mine tailings and similar soil ecosystems.

A framework for tracing timber following the Ukraine invasion

Scientific testing including stable isotope ratio analysis (SIRA) and trace element analysis (TEA) is critical for establishing plant origin, tackling deforestation and enforcing economic sanctions. Yet methods combining SIRA and TEA into robust models for origin verification and determination are lacking. Here we report a (1) large Eastern European timber reference database (Betula, Fagus, Pinus, Quercus) tailored to sanctioned products following the Ukraine invasion; (2) statistical test to verify samples against a claimed origin; (3) probabilistic model of SIRA, TEA and genus distribution data, using Gaussian processes, to determine timber harvest location. Our verification method rejects 40–60% of simulated false claims, depending on the spatial scale of the claim, and maintains a low probability of rejecting correct origin claims. Our determination method predicts harvest location within 180 to 230 km of true location. Our results showcase the power of combining data types with probabilistic modelling to identify and scrutinize timber harvest location claims.

Improved organic and pesticide-free rice (Oryza sativa L.) authentication based on multiple stable isotope ratio analysis and rice milling state

This study looked at the application of multiple bulk stable isotope ratio analysis to accurately authenticate organic rice and counteract organic fraud within the expanding global organic market. Variations of δ13C, δ15N, δ18O, and δ34S in organic, pesticide-free, and conventional rice were assessed across different milling states (brown, milled, and bran). Individual stable isotope ratio alone such as δ15N demonstrated limited capacity to correctly differentiate organic, pesticide-free, and conventional rice. A support vector machine model—incorporating δ13C, δ15N, δ18O, and δ34S in milled rice—yielded overall predictability (95%) in distinguishing organic, pesticide-free, and conventional rice, where δ18O emerged as the pivotal variable based on the feature weights in the SVM model. These findings suggest the potential of multi-isotope and advanced statistical approaches in combating organic fraud and ensuring authenticity in the food supply chain.

Stable isotope ratios and current-use pesticide levels in edible insects: Implications on chemical food safety

In the past years, the European Union (EU) has added edible insects to the list of novel foods, allowing an increasing number of insect-based products into the European market. With insects gaining more popularity in the Western world, it is crucial to investigate their chemical food safety. This study aimed at investigating possible isotopic patterns in different edible insect species (n = 52) from Asia, Africa and Europe using stable isotope ratio analysis (SIRA) to provide a framework for future investigations on food authenticity and traceability. Additionally, complementary mass-spectrometric screening approaches were applied to gain a comprehensive overview of contamination levels of current-use pesticides (CUPs) in edible insects, to assess their chemical food safety. SIRA revealed significant differences between countries in δ13CVPDB- (p < 0.001) and δ15Nair- (p < 0.001) values. While it was not possible to distinguish between individual countries using principal component analysis (PCA) and linear discriminative analysis (LDA), the latter could be used to distinguish between larger geographical areas (i.e. Africa, Europe and Asia). In general, African samples had a more distinct isotopic profile compared to European and Asian samples. When comparing the isotopic compositions of samples containing pesticides with samples with no detected pesticides, differences in sulphur compositions could be observed. Additionally, LDA was able to correctly classify the presence of pesticides in a sample with 76% correct classification based on the sulphur composition. These findings show that SIRA could be a useful tool to provide a framework for future investigations on food authenticity and traceability of edible insects. A total of 26 CUPs were detected using suspect screening and an additional 30 CUPS were quantified using target analysis, out of which 9 compounds had a detection frequency higher than 30%. Most detected pesticides were below the maximum residue levels (MRLs) for meat, suggesting low contamination levels. However, dichlorvos and fipronil could be detected in the same order of magnitude as the MRLs, even in samples purchased in Europe. These findings indicate a limited chemical risk for edible insects regarding pesticide contamination. Nevertheless, the study also highlights that further and more extensive investigations are needed to give a comprehensive assessment of the chemical risk of edible insects as a novel food source in Europe. With insects recently being potentially more incorporated into daily diets, more attention should be paid to possible chemical hazards to accurately assess their risk and to ensure food safety.

Authentication of Fennel, Star Anise, and Anise Essential Oils by Gas Chromatography (GC/MS) and Stable Isotope Ratio (GC/IRMS) Analyses.

The aromatic compound (E)-anethol is widely used in the flavor, fragrance, and medicinal industries. This compound is commonly produced through steam distillation of fennel, star anise, and anise seed. Given the cost of production, these natural and authentic essential oils are commonly adulterated with lower-cost natural materials or synthetic alternatives. The current study investigates essential oil profiles (gas chromatography/mass spectrometry) and stable isotope ratios (gas chromatography/isotope ratio mass spectrometry) of the abundant compound (E)-anethol in both authentic reference standards (n = 15) and commercially available samples (n = 30). This multifaceted analytical approach establishes techniques for ensuring the authenticity of essential oil sources of (E)-anethol and was then used to evaluate the current essential oil market sources of (E)-anethol. These findings report that adulteration of (E)-anethol-containing natural products takes various forms, and a multifaceted analytical approach is recommended for authentication. Of the commercial samples analyzed for this report, 27% were adulterated.

An optimized method for extraction and purification of inorganic phosphate from plant material for oxygen isotope ratio analysis

Compound-specific stable isotope ratio analysis of oxygen isotopes in inorganic phosphate can be used to study biological phosphorus cycling and the transformation processes controlling the fate of phosphorus. However, methods for extraction of inorganic phosphate from plant tissue for oxygen isotope ratio analysis are not consistent. Further, the purification into solid silver phosphate can be challenging and laborious. In this work, a detailed and optimized method to provide a more consistent, easily implementable and reproducible extraction using trichloroacetic acid and subsequent purification of inorganic phosphate from plant material for oxygen isotope ratio analysis is presented. Key focus points were: uniform extraction of inorganic phosphate from barley leaves, removal of dissolved organic material, flexibility in regards to the amount of inorganic phosphate extracted for the purification into silver phosphate, reduced use of chemicals and, removal of co-precipitated oxygen-bearing compounds before analysis. Most notable optimizations to the method and associated effects were:•Drying of plant material before inorganic phosphate extraction increases the method applicability to a broader range of plant sample types.•Removal of dissolved organic matter improves inorganic phosphate purification.•Sample volume adjustment according to inorganic phosphate content is vital for effective and quantitative precipitations.

Effects of sampling time and location on the geographical origin traceability of protected geographical indication (PGI) Hongyuan yak milk: Based on stable isotope ratios

Hongyuan yak milk is a protected geographical indication (PGI) product of rich nutritional value, which is popular among consumers. Stable isotope ratio analysis (SIRA) is an effective way to protect the authenticity of the geographical origin of PGI products, and it is crucial to study the factors affecting stable isotopes. Firstly, we proved that the SIRA could be used to identify the geographical origin of Hongyuan yak milk, and that the identification accuracy in combination with δ13C and δ18O was 100 %. Secondly, we analyzed the effect of sampling selection on the stable isotopes of Hongyuan yak milk in practical applications, which showed that sampling time influenced the δ13C, δ2H, and δ18O, while the sampling locations did not. There were interactions between the effect of sampling time and location on δ2H and δ18O. These results provide a reliable method for identifying PGI products and also provide new guidance on sampling models.

Early Neolithic pastoral land use at Alsónyék-Bátaszék, Hungary (Starčevo culture): New insights from stable isotope ratios

The earliest introduction of livestock (cattle, goats, sheep, pigs) into the Carpathian Basin was an important step towards farming expansion into continental Europe. This spread beyond the environments of the southern Balkans was accompanied by a reduction in the spectrum of cultivated crops, changes in the relative representation of different domestic animals, and, most likely, adaptations of husbandry practices. How the earliest farmers in the Carpathian Basin kept their domestic stock is still understudied. We explored early animal management and land use strategies at the Starčevo settlement at Alsónyék-Bátaszék, Hungary (Early Neolithic, ca. 5800–5600 cal BC). Settled at the intersection of wide alluvial plains, waterlogged meadows and marshes to the east, and forested hills to the west, early farmers at Alsónyék had a wide variety of options for nourishing their livestock. We performed stable isotope ratio analysis of bone collagen (n = 99; δ13C, δ15N) and tooth enamel (nteeth = 28, sequentially sampled for δ13C and δ18O) from wild and domestic animals to locate them in the landscape and investigate herding practices on a seasonal scale. The bone collagen isotope ratios mostly indicate feeding in open environments. However, results from the sequential analysis of cattle and sheep enamel suggest diverse dietary strategies for winters, including consumption of forest resources, consumption of summer hay and grazing in an open environment. Most pigs appear to have had herbivorous diets, but several individuals likely supplemented their diet with animal protein. Stable isotope ratio results from the Lengyel phase at Alsónyék (ca. 4800–4300 cal BC) suggest more access to animal protein for pigs, and feeding in more open areas by wild boar, red deer and cattle compared to the Starčevo phase. This study’s results demonstrate considerable variability in early animal husbandry practices at Alsónyék.

The influence of taphonomy on histological and isotopic analyses of treated and untreated buried modern human bone

The chemical (e.g., preservation/embalming) treatment of skeletal remains can reduce overall DNA quality and quantity. The histological and stable isotope examination of treated and untreated human remains improves our understanding of how chemical preservatives impact bone diagenesis and will determine if chemical treatment adversely affects stable isotope ratio analysis of collagen. Fidelity in the application(s) of stable isotope interpretations requires that the isotope delta (δ) values have not been altered postmortem. Re-associated antimeres and refits of chemically treated and untreated rib and long bones from eight casualties [thin-sectioned human bone (n = 43) and collagen extraction/stable isotope analysis (n = 42)] from the World War II Battle of Tarawa were examined to compare skeletal elements from the same individual that had different taphonomic histories. Histological analyses included scoring upon the Oxford Histological Index (OHI) and Birefringence scale, recording microbial invasion, and general observations. The collected data were analyzed via simple descriptive statistics and paired samples t-tests. Treated remains scored higher on the OHI and for Birefringence, indicating that bone quality was good to excellent. The untreated samples scored lower on the OHI and Birefringence scales suggesting poorer preservation than the treated remains. Histology results were supported by the isotope sample preparation results: the collagen % yield was higher for treated bone than untreated bone. Additionally, chemical preservation had no meaningful impact on isotope δ values of treated and untreated remains from the same element or pair-matched elements. Overall, treated remains exhibited good preservation while untreated remains exhibit poorer preservation with significant microfocal destruction to the extent that little histological analyses can be applied. Stable isotope ratio analysis is viable for both treated and untreated remains indicating this testing modality likely can be used for most treated remains, regardless of origin.

Verifying origin claims on dairy products using stable isotope ratio analysis and random forest classification.

Scientifically underpinning geographic origin claims will improve consumer trust in food labels. Stable isotope ratio analysis (SIRA) is an analytical technique that supports origin verification of food products based on naturally occurring differences in isotopic compositions. SIRA of five relevant elements (C, H, N, O, S) was conducted on casein isolated from butter (n=60), cheese (n=96), and whole milk powder (WMP) (n=41). Samples were divided into four geographic regions based on their commercial origin: Ireland (n=79), Europe (n=67), Australasia (n=29) and USA (n=22). A random forest machine learning model built using δ13C, δ2H, δ15N, δ18O and δ34S values of all products (n=197) accurately (88% model accuracy rate) predicted the region of origin with class accuracy of 95% for Irish, 84% for European, 71% for Australasia, and 94% for US products.

Using stable carbon isotope ratio analysis to detect adulteration in red yeast rice dietary supplements

Red yeast rice (RYR) is marketed as a dietary supplement because it contains natural 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins), including monacolin K. However, there is concern that some RYR supplements may be adulterated with the pharmaceutical drug lovastatin to enhance health claims. We have developed an optimized method to isolate monacolin K/lovastatin from complex RYR dietary supplement matrices to then test for adulteration in RYR supplements using stable carbon isotope (δ13C) analysis. Samples were initially screened for monacolin K/lovastatin using liquid chromatography with mass spectrometric detection (LC-MS). To ensure the extraction process did not affect the measured isotopic values (i.e., isotopic fractionation effects), neat lovastatin standards were spiked into two types of blank RYR matrices (powder and gel). The monacolin K/lovastatin peaks were detected using high performance liquid chromatography with ultraviolet detection (HPLC-UV) and isolated using fraction collection. Residual matrix components were removed from targeted fractions by solid phase extraction (SPE) using graphitized carbon black cartridges. The resulting isolates were then analyzed using elemental analyzer-isotope ratio mass spectrometry (EA-IRMS) to measure δ13C values. The δ13C values of the extracted lovastatin standards were compared to their respective neat lovastatin δ13C values and demonstrated negligible isotopic fractionation effects. Using this optimized clean up method and carbon isotope analysis, thirty-one samples were screened. Eight RYR dietary supplement samples had >0.8 mg/g of monacolin K/lovastatin, our minimum threshold for analyzing samples using this method. Four of these eight samples had δ13C values greater than −28.3‰, a previously proposed cutoff value for natural monacolin K, indicating likely adulteration. Additionally, five RYR powder samples were analyzed as part of a collaborative study using in-house methods from two laboratories and the data shows acceptable agreement in the δ13C values of monacolin K/lovastatin (differences ranging from ±0.02‰ to ±0.76‰). This optimized method represents a robust, reproducible procedure for detecting lovastatin adulteration in dietary supplements with minimal isotopic fractionation.

A dietary perspective of cat-human interactions in two medieval harbors in Iran and Oman revealed through stable isotope analysis

Cats are hypercarnivorous, opportunistic animals that have adjusted to anthropogenic environments since the Neolithic period. Through humans, either by direct feeding and/or scavenging on food scraps, the diet of cats has been enriched with animals that they cannot kill themselves (e.g., large mammals, fish). Here, we conducted carbon and nitrogen stable isotope ratio analysis to reconstruct the diet of medieval cats and investigate cat-human interactions in two medieval harbor sites (Qalhât, Oman and Siraf, Iran). The analysis included 28 cat individuals and 100 associated marine and terrestrial faunal samples pertaining to > 30 taxa. The isotopic results indicate a high marine protein-based diet for the cats from Qalhât and a mixed marine-terrestrial (C4) diet for the cats from Siraf. Cats at these sites most likely scavenged on both human food scraps and refuse related to fishing activities, with differences in the two sites most likely associated with the availability of marine resources and/or the living conditions of the cats. By shedding light on the dietary habits of cats from two medieval harbors in the Arabian Gulf and Gulf of Oman, this study illustrates the potential of stable isotope analysis in reconstructing human-cat interactions in the past.

Bulk and fatty acid-specific stable isotope analysis coupled with multivariate modeling for differentiating free-range grass-fed and captive grain-fed yak meat

The husbandry pattern of yak meat is a crucial factor for its market price and competitiveness; deliberately mislabeling of the production information will cause consumer confidence crisis. In this work, stable isotope signatures of free-range grass-fed (FRG) and captive grain-fed (CG) yak meat were analyzed for their discrimination. δ13C, δ2H, δ18O, δ15N, δ34S of bulk muscle and δ13C of six fatty acids were respectively determined by EA–IRMS and GC–C–IRMS. ANOVA and correlation analysis of stable isotope ratios of yak muscle and fatty acids against its feed indicated that their δ13C, δ15N, δ34S significantly are related, which provides the evidence of diet and trophic information. δ13C of six fatty acids (FAs) are significantly different (p < 0.05) between two patterns; all FAs of FRG yak meat are highly related to grass silage (R > 0.70), but saturated and unsaturated FAs of CG yak meat are respectively related to soybean silage (R > 0.70) and maize silage (R > 0.56). PLS-DA and LDA modeling based on stable isotope ratios of yak muscle and FAs together discriminate yak meat from these two husbandry patterns with accuracies of 100 %. This strategy may be promising as a feasible method for confirming the husbandry patterns of yak meat, ensuring label authenticity and food safety.